1.

INTRODUCTION
1.1Background--------------------------------------------------------------------------------------------------- 1
1.2.1 Classification of amylase---------------------------------------------------------------------- 2
1.2.1.2 β-amylase------------------------------------------------------------------------------------------2
1.2.1.3 γ-amylase-----------------------------------------------------------------------------------2
1.3 Starch----------------------------------------------------------------------------------------------------------3
1.4 Application of amylases------------------------------------------------------------------------------------5
1.4.1 Starch hydrolysis and fructose production--------------------------------------------------5
1.4.2 Manufacturing of maltose---------------------------------------------------------------------------5
1.4.3 Production of anomalously linked oligosaccharides mixture (Alo mixture) -----------6
1.4.4 Detergents
1.4.5 Paper and pulp industry
1.4.6 Baking
1.4.7 Removal of starch sizer from textile (desizing)
1.4.8 Direct fermentation of starch to ethanol
1.4.9 Manufacture of maltotetraose syrup
1.4.10 Manufacture of oligosaccharides mixture
1.4.11 Manufacture of high fructose containing syrups
1.4.12 Treatment of starch processing waste water (SPW)

2. LITERATURE REVIEW
2.1 Background
2.2 Microbial Sources of Amylase
2.3 Purification and Characterisation of Α-Amylase
2.4 Effect of Temperature and pH
2.5 Effect of Metal Ions
2.6 Effect of Additives

3. AIMS AND OBJECTIVES

3.1 Aim
3.2 Objectives

4. MATERIALS AND METHOD

4.1 Sample collection
4.2 Total viable count
4.3. Isolation and purification of bacterial strain
4.3.1 Enrichment
 4.3.2. Isolation
4.3.3 .Purification of isolate
4.4. Screening of the amylase producing bacterial strain
4.4.1 .Enrichment of amylase producer
4.4.2. Isolation of amylase producer
4.5. Identification of amylase producer isolate
4.6. Optimization of culture condition
4.6.1. Effect of incubation period
4.6.2. Effect of Carbon Sources on the Production of Extracellular Amylases
4.6.3. Determination of optimium concentration of starch on the Production of
Extracellular amylases
4.6.4. Effect of Nitrogen Sources on the Production of Extracellular Amylases
4.6.5 Determination of optimum concentration of peptone on the production of
extracellular amylase
4.6.6. Determination of optimum temperature on the production of extracellular
amylases
4.6.7. Determination of optimum pH on the Production of Extracellular amylases
4.7. Production and extraction of crude extracellular amylase
4.8. Partial purification of crude extracellular amylase
4.8.1 Ammonium Sulphate Precipitation:

4.8.2. Dialysis
4.9 Protein Estimation
4.10. Amylase assay
4.10.1 Standard maltose curve
4.11. Effect of pH on the partially purified amylase
4.12. Effect of temperature of the partially purified amylase
4.13. Effect of various metal ions on partially purified amylase
4.14. Effect of oxidants and surfactants on partially purified amylase
4.15. Statistical analysis

5. RESULTS
5.1. Total viable count
5.2. Isolation, purification and screening of amylase producer:
5.3 Identification of amylase producing isolate
5.4 Optimization of culture condition
5.4.1 Determination of optimum incubation period:
5.4.2 Determination of optimum temperature
5.4.3 Determination of optimum pH
5.4.4 Determination effect of different carbon source
5.4.6 Determination of optimum concentration of different starch concentration
5.4.7 Determination of effect of different nitrogen concentration
5.4.8 Determination of optimum peptone concentration
5.4.9 Determination of effect of different NaCl concentration
5.5. Partial purification of amylase:
5.6 Characterization of partially purified amylase from Bacillus sp. AY-27
 5.6.1 Determination of effect of temperature on amylase activity
5.2.2 Determination of effect of pH on amylase activity
5.2.3 Determination of effect metal ions and additives

6. DISCUSSION

7. SUMMARY AND CONCLUSION

BIBLIOGRAPHY

APPENDIX